Biliprotein light-harvesting strategies, phycoerythrin 566.
A series of experiments on the light-harvesting properties of the cryptomonad biliprotein phycoerythrin 566 has been carried out on purified protein isolated from Cryptomonas ovata. Although this pigment has an absorption maximum at 566 nm, a property very close to that of other phycoerythrins, it was found to have a totally unique set of chromophores. The chromophores (bilins) responsible for its absorption spectrum were analyzed by a number of approaches. Chromophore-containing peptides were produced by trypsin treatment and purified in order to isolate the individual peptide-bound bilins free of overlapping absorption. These chromopeptides, after comparison with appropriate controls, showed that three spectrally distinct bilins occurred on the purified oligomeric protein. Two of the bilins were the well-known phycoerythrobilin and cryptoviolin, but the third was previously undiscovered and had an absorption spectrum between that of cryptoviolin and phycocyanobilin. Since the spectral diversity of the three bilins was fully maintained in solvents that minimize the effects of apoprotein on the spectra of the bilins, it is likely that the three bilins are also structurally dissimilar. The alpha and beta subunits, which constitute the protein, were separated by ion-exchange chromatography, and the new bilin was found to be the sole chromophore on the alpha subunit. It was also found that at least two alpha subunits could be separated and they both had this unusual bilin (cryptobilin 596). The beta subunit, therefore, contained both phycoerythrobilin and cryptoviolin. On the basis of the spectra of the three chromopeptides, the absorption spectrum of the protein was modeled using the known absorptivities of cryptoviolin and phycoerythrobilin.
MacColl
R
R
Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany 12201-0509.
Guard-Friar
D
D
Ryan
T J
TJ
D000818:Animals; D001654:Bile Pigments / Q000032:analysis; D056890:Eukaryota / Q000032:analysis*; D045342:Light-Harvesting Protein Complexes; D010446:Peptide Fragments / Q000302:isolation & purification; D052980:Phycobilins; D010798:Phycocyanin; D010799:Phycoerythrin / Q000302:isolation & purification; D010860:Pigments, Biological / Q000302:isolation & purification; D010940:Plant Proteins / Q000302:isolation & purification; D011758:Pyrroles / Q000302:isolation & purification; D013053:Spectrophotometry; D045725:Tetrapyrroles; D014357:Trypsin
D001654:Bile Pigments; D045342:Light-Harvesting Protein Complexes; D010446:Peptide Fragments; D052980:Phycobilins; D010860:Pigments, Biological; D010940:Plant Proteins; D011758:Pyrroles; D045725:Tetrapyrroles; D010798:Phycocyanin; D010799:Phycoerythrin; C011070:phycoerythrobilin; D014357:Trypsin